Volume 13, Issue 2, March – April 2012; Article-008 ISSN 0976 – 044X
R. Sathish
1
, R. Murugesan
2
, Arul Amuthan
3*
1
Lecturer, Sri Sairam Siddha Medical College, Chennai, India.
2
Scientific Officer, Sophisticated Analytical Instrument Facility, Indian Institute of Technology Madras, India.
3
Lecturer in Pharmacology, Melaka Manipal Medical College, Manipal University, India.
*Corresponding author’s E-mail: dramuthanmd@yahoo.co.in
Accepted on: 16-01-2012; Finalized on: 20-03-2012.
ABSTRACT
Siddha system of medicine believes herbometallic formulation to be potent than herbal formulation alone. So far, fewer studies have been conducted on standardization of such preparations. Mega sanjeevi mathirai (MSM), a traditional Siddha herbometallic drug was prepared as per the procedures mentioned in a siddha literature. The chemical finger print was taken by using modern analytical techniques like Inductive coupled plasma optical emission spectroscopy (ICP-OES) and Fourier transform infra-red spectroscopy (FTIR). In addition, the particle size of mega sanjeevi mathirai was also assessed by High Resolution Scanning Electron
Microscope (HR SEM). The results confirmed the absence of lead, arsenic and cadmium. Reports showed the presence of inorganic elements such as mercury (2.96ppm), sodium (6.26ppm), potassium (4.22ppm), calcium (15.10ppm), phosphorous (3.63ppm) and sulphur (6.25ppm). FTIR confirmed the presence of organic moieties as follows; IR (KBr, cm
-1
), 3399 (-OH structure of –COOH group),
1716 (C=O structure of carboxylic acid), 1618 (C=N structure), 1214 (Asymmetric C-O-C structure), 1047 (Symmetric C-O-C structure).
The range of particle size varies between 3 10 µ. These findings could be used as a chemical fingerprint for future referen ce of
Mega sanjeevi mathirai in chemical standardization.
Keywords: Metallic Drug, Ayurveda, Basma, Traditional Medicine, Drug Standardization, Mercury.
INTRODUCTION
Indian systems of medicines have been widely used for thousands of years in India. Siddha system is the ancient and unique among the Indian system of medicines.
Standardization of traditional drug is a burning topic in today’s drug research industry. Standardization in herbal formulation is difficult, because they are usually mixtures of many constituents and the active principle in most cases is unknown
1
. In Siddha system of medicine, the drug sources are obtained from plants, inorganic ores and animals.
Gradual unavailability of herbals makes attention towards utilization of metal based drugs and animal based drugs for therapeutic purpose. Now days, acceptance of traditional system of medicine in developed countries is sharply increasing
2
. Proper standardization of drug preparation methods as well as chemical analysis of traditional formulation is mandatory to gain support for its use worldwide. Very few studies have been done on the chemical standardization, efficacy and safety aspects of herbo-metallic traditional drugs.
MATERIALS AND METHODS
Preparation of mega sanjeevi mathirai
The drug was prepared as per the standard procedures mentioned in Siddha formulary literature Anupoga vaithiya navaneetham. All the ingredients were bought from authorized Siddha raw material shop in Chennai,
India. Lingam (cinnabar – red sulphide of mercury), Rasa chenduram (mercury sulphide), Veeram (mercury perchloride) and Pooram (mercury subchloride) were taken in the quantity of 8.75g each. All the four inorganic raw materials were further processed individually with selective liquids for purification process (Suddhi muraigal). The above four raw inorganic materials were either dipped in the liquids or boiled with the liquids according to the procedure. The liquids added for the purification process include honey, cow milk, lemon fruit juice, Acalypha indica leaf juice, Mukiya maderaspatna leaf juice and Piper nigram seed decoction. After the purification process, above said four inorganic materials were mixed with 35g pericarp powder of Terminalia
chebula dry fruit and grinded continuously for 15hr by adding 300ml of lime juice. Then, grinded outcome was made as pills of 65mg, which was dried in room temperature
3
(Figure 1).
Mega sanjeevi mathirai (MSM), a herbo metallic Siddha drug is mainly used to treat infectious genital diseases, sexually transmitted diseases, chronic urinary tract infection and cystitis especially in chronic diabetes mellitus patient
3
.
Since this drug was not chemically standardized, the aim of our present study was to standardize the chemical composition of the mega sanjeevi mathirai by modern analytical techniques. Inorganic elements of all four raw materials were quantified by Inductive coupled plasma optical emission spectroscopy (Optima 5300 DV ICP-OES) equipped with a
Sea Spray concentric nebulizer (Glass Expansion,
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Estimation of inorganic elements and Infrared spectra in raw materials

Volume 13, Issue 2, March – April 2012; Article-008 ISSN 0976 – 044X
Pocasset, MA) and cyclonic spray chamber. Following procedures were followed: nebulizer flow, 0.8/min ; radiofrequency power, 1450 W; sample introduction, 1.5 ml min
-1
; flush time, 20 s; delay time, 10 s; read time, 10 s; wash time, 30 s; and replicates, three. Standards were prepared by dilution of 1000 mg l
-1
stock solutions and the calibration curve was obtained using five to ten points including the blank.
-1
Fine powder of MSM was analyzed for particle morphology by High Resolution Scanning Electron
Microscope (JEOL ASM 3500 HR-SEM). A representative portion of each sample was sprinkled onto a double side carbon tape and mounted on aluminum stubs in order to get a higher quality secondary electron image for SEM examination
1
.
RESULTS AND DISCUSSION
Raw materials were powdered and the infrared spectral characterization was obtained by using Perkin-Elmer FTIR
Spectrophotometer in the region (4000-450 cm-1) by KBr pellet method
1
.
Changes of inorganic element composition in purification process in raw materials
Toxic metals like arsenic, cadmium and lead were absent in all the raw materials. Mercury level has been drastically reduced in the purified raw materials. Thus, the intention of purification process of metallic raw materials by using herbal juices could to reduce the toxic metal quantity in the herbo-metallic formulations. All other elements have been considerably reduced after purification process except sulphur in lingam. (Table 1).
Changes of Infra-red spectra after purification process in raw materials
No major changes were observed in spectra of lingam and rasachenduram, which showed almost similar spectra before and after purification process (Figure 2, 3, 4 & 5).
ICP-OES and FTIR were repeated after the purification processes in the purified raw materials was done in order to assess the chemical modulations because of purification processes.
Estimation of inorganic elements, Infrared spectra and drug particle size in MSM
Figure 1: Flow chart of drug preparation procedure of
Mega sanjeevi mathirai
Estimation of inorganic elements and Infrared spectra in purified raw materials
In purified veeram, acidic (OH) (3361) moiety and CO moiety of carboxylic acid (1715) were introduced.
Addition of these functional groups might be the purpose for the purification process in veeram (Figure 6 & 7).
Purification procedure of pooram leads to the insertion of numerous moieties. Ester group (1733) has been converted in to acidic (1716) moiety. Hydroxyl (OH) group
(3503) has been converted into carboxylic (COOH) moiety
(3349). Other notable insertion after purification of pooram include aromatic, CH2, CH3 and ether (-COC) link
(1064) (Figure 8 & 9). These findings have given the clue that “Suddhi muraigal” in the traditional drug preparation has some significance in chemical structural modulation.
After the mega sanjeevi mathirai was prepared, ICP-OES and FTIR were repeated to assess the final chemical composition in the product.
Table 1: Analysis of inorganic elements by ICP-OES in raw materials before & after purification process and in final product mega sanjeevi mathirai
Raw material name lingam (raw)
Lead
BDL
Cadmium
BDL
Inorganic elements in parts per million (ppm)
Arsenic
BDL
Mercury
113.39
Sulphur
114.2
Na
+
K
+
Ca
2+
Phosphorus
1.33 207.33 25.72 14.23
Lingam (after purification)
Rasacehnduram (raw)
Rasachenduram (after purification)
Veeram (Raw)
Veeram (after purification)
Pooram (raw)
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
Pooram (after purification) BDL BDL
MSM BDL BDL
BDL - below detectable level; MSM – Mega sanjeevi mathrai
BDL
BDL
BDL
BDL
BDL
BDL
BDL
BDL
70.83
201.4
123.3
117.7
80.59
241.5
100.23
2.96
217.3
258.9
207.9
7.23
2.29
14.21
10.51
6.25
1.22
9.51
5.20
8.83
6.73
3.29
1.24
6.26
17.13
22.15
19.14
27.05
17.68
17.13
12.16
4.22
2.28
28.61
18.30
12.08
2.88
23.70
20.26
15.10
1.1
9.51
5.1
6.1
4.12
8.12
7.16
3.63
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Volume 13, Issue 2, March – April 2012; Article-008 ISSN 0976 – 044X
Figure 2: Infra-red spectra of lingam before purification process
Figure7: Infra-red spectra of veeram after purification process
Figure 3: Infra-red spectra of lingam after purification process
Figure 8: Infra-red spectra of pooram before purification process
Figure 4: Infra-red spectra of rasachenduram before purification process
Figure 9: Infra-red spectra of pooram after purification process
Figure 5: Infra-red spectra of rasachenduram after purification process Figure 10: Infra-red spectra of mega sanjeevi mathirai
Chemical composition of Mega sanjeevi mathirai
Figure 6: Infra-red spectra of veeram before purification process
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ICP-OES revealed the absence of toxic metals like lead, cadmium and arsenic. But amount of mercury presented in this drug was 2.96ppm, which is above the permitted limit (1 ppm)
4
. High mercurial amount in this drug alarms for further study on toxicity profile. Detailed study is necessary to evaluate whether the herbal components have any protective role or chelation role in mercury toxicity. Other inorganic elements like sodium, potassium,

Volume 13, Issue 2, March – April 2012; Article-008 ISSN 0976 – 044X calcium, phosphorous and sulphur were also present in minimal quantity (Table 1).
Infra-red spectra of mega sanjeevi mathirai have shown few peaks as follows;
3399 (-OH structure of –COOH group), 1716 (C=O structure of carboxylic acid), 1618 (C=N structure), 1214
(Asymmetric C-O-C structure) and 1047 (Symmetric C-O-C structure). These organic moieties might be responsible for the therapeutic values (Figure 10).
HR SEM analysis revealed that the particle size of MSM is in the range of 3 to 10µ. Shape of the particles was cubical and the surface was smooth (Figure 11). acid, C=N structure, asymmetric C-O-C structure and
Symmetric C-O-C structure were also present in mega sanjeevi mathirai. This report could be used as chemical finger print for future references in chemical standardization of Mega sanjeevi mathirai.
Further detailed studies are required to evaluate the importance of “Suddhi muraigal” in Siddha drug preparation technique, which may reveal the scope for chemical modulation by traditional methods in pharmaceutical industry. Since the chemical composition is mainly depend on the herbals, which might contain varied amount of phytochemicals in response to different climate or different geological location, similar studies have to be repeated by using herbals collected from different locations and the variations in chemical composition have to be further documented.
Figure 11: HR SEM photo of Mega sanjeevi mathirai
CONCLUSION
In this study, Mega sanjeevi mathirai was prepared and analyzed according to the standard procedures. There were notable changes in spectra after the suddhai muraigal (purification process). The findings in Mega sanjeevi mathirai revealed the absence of heavy metals like lead, arsenic and cadmium. But mercury was present above the permitted level. Organic moieties like OH structure of –COOH group, C=O structure of carboxylic
About Corresponding Author: Dr. Arul Amuthan
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Sayabu HA. Anupoga Vaithiya Navaneetham Vol.VII
(Tamil). India:Thamatai publication; 2002. P.74-5.
4.
Lohar DR. Protocol For Testing Ayurvedic, Siddha &
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Dr. Arul Amuthan is an ethnopharmacologist, graduated in Siddha Medicine (Indian Traditional
Medicine) from Tamilnadu Dr.MGR Medical university and post graduated in Pharmacology from Manipal University. He was awarded by Jawaharlal Nehru Memorial Fund for securing university first in his post-graduation. He is doing intense research in traditional medical systems in the area of drug standardization, drug discovery, pharmacological and toxicological evaluation.
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